EVAPORATION DURING PREPARATION OF UNSUPPORTED THIN VITRIFIED AQUEOUS LAYERS FOR CRYOELECTRON MICROSCOPY
EVAPORATION DURING PREPARATION OF UNSUPPORTED THIN VITRIFIED AQUEOUS LAYERS FOR CRYOELECTRON MICROSCOPY
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DOI：
10.1002/jemt.1060160407
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发表时间：
1990-12-01
期刊：
影响因子：
--
通讯作者：
DUBOCHET, J
中科院分区：
文献类型：
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作者：
CYRKLAFF, M;ADRIAN, M;DUBOCHET, J
当制备用于低温电子显微镜检查的无支撑水性悬浮液的薄玻璃化膜时，水的蒸发不能完全避免。这导致溶质浓度增加，这可能影响观察到的材料。我们已经定量研究了这种效果，通过测量的对比度的聚苯乙烯球甲泛葡胺溶液。当在亲水性多孔支持物上制备样品时，干燥效应通常可以忽略不计，但当使用疏水膜时，干燥效应通常很重要。湿空气流、使薄液膜最少暴露于大气的双重吸墨或优化吸墨条件的自动柱塞都是减少干燥效应的简单方法。利用作用于亲水性支撑膜的该第三装置，溶质浓度的增加被限制为小于20%。
Evaporation of water cannot be fully avoided when an unsupported thin vitrified film of an aqueous suspension is prepared for cryo‐electron microscopy. This results in increasing concentration of solute which could affect the observed material. We have quantitatively studied this effect by measuring the contrast of polystyrene spheres in a metrizamide solution. The drying effect is generally negligible when specimens are prepared on a hydrophilic perforated support but it is frequently important when hydrophobic films are used instead. A flow of humid air, double blotting with minimal exposure of the thin liquid film to the atmosphere, or an automatic plunger optimizing the blotting conditions are simple methods for reducing drying effects. With this third device acting on a hydrophilic supporting film, the increase of solute concentration is limited to less than 20%.